2.The interaction between secreted frizzled-related protein 2 and osteoblast-specific factor 2 in keloid.
Zhen-Xiang WANG ; Xi TAO ; Shi-Rong LI
Chinese Journal of Burns 2009;25(2):126-128
OBJECTIVETo verify the interaction between secreted frizzled-related protein 2 (SFRP2) and osteoblast-specific factor 2 (OSF-2).
METHODSHA-tagged OSF-2 fusion protein recombinant vector pCMV-HA-OSF-2, which could express in mammal cells was constructed, then identified by enzyme-cutting and transfected into human kidney 293 (HK293) cells with or without Myc-SFRP2 recombinant eukaryotic expression vector pCMV-HA-SFRP2. The interaction between SFRP2 and OSF-2 was detected through coimmunoprecipitation and Western blotting.
RESULTSIn electrophoresis bath, target fragment of SFRP2 coding gene with 800 bp and target gene OSF-2 with 2500 bp could be seen respectively after enzyme-cutting, which showed that pCMV-Myc-SFRP2 and pCMV-HA-OSF-2 were constructed successfully. No HA-OSF-2 expression was detected after pCMV-Myc-SFRP2 or pCMV-HA-OSF-2 transfection. Whereas, HA-OSF-2 expressed by Myc antibody immunoprecipitation after pCMV-Myc-SFRP2 and pCMV-HA-OSF-2 co-transfection.
CONCLUSIONSHA-OSF-2 recombinant vector can express in mammal cells. Interaction exists between HA-OSF-2 and SFRP2.
Cell Line ; Core Binding Factor Alpha 1 Subunit ; genetics ; metabolism ; Genetic Vectors ; Humans ; Keloid ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Recombinant Fusion Proteins ; metabolism ; Transfection
4.Development and evaluation of immunoassay for zeranol in bovine urine.
Yuan LIU ; Cun-zhen ZHANG ; Xiang-yang YU ; Zhi-yong ZHANG ; Xiao ZHANG ; Rong-rong LIU ; Xian-jin LIU ; Zhen-ming GONG
Journal of Zhejiang University. Science. B 2007;8(12):900-905
A high affinity polyclonal antibody-based enzyme linked immunosorbent assay (ELISA) was developed for the quantification of zeranol in bovine urine. On the basis of urine matrix studies, the optimized dilution factors producing insignificant matrix interference were selected as 1:5 in pretreatment. In the improved ELISA, the linear response range was between 0.02 and 1 microg/ml, and the detection limit was 0.02 microg/ml for the assay. The overall recoveries and the coefficients of variation (CVs) were in the range of 82% to approximately 127% and 3.5% to approximately 8.8%, respectively. Thirty-six bovine urine samples spiked with zeranol (ranging from 0.2 to 10 microg/ml) were detected by the ELISA and liquid chromatography (LC) method, and good correlations were obtained between the two methods (R(2)=0.9643). We conclude that this improved ELISA is suitable tool for a mass zeranol screening and can be an alternative for the conventional LC method for zeranol in bovine urine.
Animals
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Calibration
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Cattle
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Chromatography, High Pressure Liquid
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Enzyme-Linked Immunosorbent Assay
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methods
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Indicator Dilution Techniques
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Zeranol
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immunology
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urine
5.Research progress on determination of lignans from Schiandra chinensis and its preparations.
Liu-qing YANG ; Xiang-yang WU ; Zuo-qi XU ; Hui-rong HOU ; Hai-zhen FU
China Journal of Chinese Materia Medica 2005;30(9):650-653
The latest research progress on quantitative determination methods of main active components-lignans from Schisandra chinensis and its preparations has been summarized, such as spectrophotometry, thin-layer chromatography scanning, high performance liquid chromatograpy, gas chromatography-mass spectrometry and capillary electrochromatography. The characteristics and application areas of every analytical method have also been stated. It offers reference on quality control of crude drug and its preparations of S. chinensis.
Capsules
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Chromatography, High Pressure Liquid
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methods
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Chromatography, Thin Layer
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methods
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Drugs, Chinese Herbal
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administration & dosage
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chemistry
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Fruit
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chemistry
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Gas Chromatography-Mass Spectrometry
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Lignans
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analysis
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Plants, Medicinal
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chemistry
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Quality Control
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Schisandra
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chemistry
6.Hypoglycemic effects of sodium metavanadate in diabetic mice and its effect on glucose phosphorylation.
Ming-zhi XU ; Ai-zhen ZHANG ; Xiang-rong LI ; Wei XU ; Ling-wei SHEN
Chinese Journal of Preventive Medicine 2003;37(3):174-177
OBJECTIVETo investigate the effects of sodium metavanadate (SMV) on blood sugar and glucose phosphorylation in mice, and to discuss the possible mechanism of its hypoglycemic effects.
METHODSDiabetic mice (D) and control mice (V) were randomly allocated to drink SMV (0.2 mg/ml) (CV and DV groups) or NaCl (80 mmol/L) (C and V groups) respectively. The study lasted for 5 weeks. Liver glucokinase, muscle hexokinase, blood glucose and insulin were assayed at the end of each week.
RESULTSBlood glucose was higher in the diabetic groups before the administration of SMV, and the blood glucose level of group DV decreased from (18.77 +/- 1.28) to (8.94 +/- 0.94) mmol/L (P < 0.01) after oral administration of SMV for one week. While liver glucokinase increased from (1.29 +/- 0.64) to (15.36 +/- 1.57) mIU/min/mg protein and muscle hexokinase increased from (1.93 +/- 0.50) to (18.62 +/- 1.71) mIU/min/mg protein (P < 0.01) respectively. There was no continuous change of these parameters during the later weeks. No significant change of serum insulin was observed in the diabetic mice. There was a remarkable negative correlation of blood glucose level with liver glucokinase and muscle hexokinase levels.
CONCLUSIONThe hypoglycemic effects of SMV was independent of insulin level. In consideration of the close relations of the activities of liver glucokinase and muscle hexokinase with diabetes, and the improving of impaired glucose phosphorylation in diabetic mice by oral sodium metavanadate, which might be the mechanism of hypoglycemic effects of SMV.
Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Experimental ; blood ; Female ; Glucokinase ; metabolism ; Hexokinase ; metabolism ; Hypoglycemic Agents ; pharmacology ; Insulin ; blood ; Liver ; metabolism ; Mice ; Mice, Inbred ICR ; Muscle, Skeletal ; metabolism ; Phosphorylation ; Random Allocation ; Vanadates ; pharmacology
7.Influence of substance P on the release of histamine in the human hypertrophic scar tissue.
Liang CHEN ; Sha LIU ; Shi-rong LI ; Lin CONG ; Ju-long WU ; Zhen-xiang WANG
Chinese Journal of Burns 2006;22(3):192-194
OBJECTIVETo investigate the influence of substance P (SP) on the release of histamine in the human hypertrophic scar tissue, and to explore the prerequisite of their interaction.
METHODSTissue specimens of normal skin and hypertrophic scar from eight hospitalized patients were excised and cut into 0.5 to 1.0 mm3 pieces, and the histamine release by mast cell (MC) under the stimulation of different concentration of SP and calcium, as well as the different affect time of SP, were determined with fluorescence spectrometer. Then the histamine release rate was calculated.
RESULTSThere was obvious release of histamine when SP concentration was 1 x 10(-6) mol/L , and the release rate was (50.0 +/- 3.6) %, which was significantly higher than that by SP in the concentration of 0 mol/L [(44.0 +/- 3.2) %, P < 0.01]. Therefore it seemed to be dose-dependent. About 90% of histamine was released within 15 minutes of 5 x 10(-1) mol/L Substance P stimulation, and it was also time-dependent. The histamine release reached the peak when calcium concentration was 5 x 10(-3) mol/L, which seemed to be dose-dependent, but it decreased transiently when calcium concentration was 1 x 10(-3) mol/L. In all occasions, the influence of SP on the histamine release by MC in hypertrophic scar (HS) was markedly higher than that in normal skin (NS) (P < 0.01). Conclusion The influence of SP on the histamine release by MC in HS was markedly higher than that in NS, and it might be closely related to itching sensation and the formation of hypertrophic scar.
Child ; Child, Preschool ; Cicatrix, Hypertrophic ; metabolism ; Female ; Histamine ; metabolism ; Humans ; Male ; Skin ; metabolism ; Substance P ; pharmacology
8.Expression of Smo protein and the downstream transcription factor Gli1 protein in Sonic hedgehog signal transduction pathway in gastric carcinoma.
Zhen-xiang RONG ; Chi-hua FANG ; Da-jian ZHU ; Sheng-jun LIU
Journal of Southern Medical University 2006;26(12):1728-1731
OBJECTIVETo study the expression of Smo protein and the downstream transcription factor Gli1 protein in Sonic hedgehog signal transduction pathway in gastric carcinoma.
METHODSA tissue microarray was constructed using 85 gastric carcinoma and 25 normal gastric mucosa specimens. The expression of Smo and Gli1 proteins were detected immunohistochemically and the correlation between their expression in gastric carcinoma was analyzed.
RESULTSOnly weak expression, if any, of Smo and Gli1 proteins was detected in normal gastric mucosa, but in papillary adenocarcinoma, tubular adenocarcinoma and poorly differentiated adenocarcinoma, their expressions were significant increased as the differentiation degree was lowered. Smo protein expression in gastric carcinoma was significantly correlated with that of Gli1 protein with correlation coefficient of 0.989 (P<0.001).
CONCLUSIONThe abnormal activity of Sonic hedgehog signal transduction pathway may play an important role in the occurrence of papillary adenocarcinoma, tubular adenocarcinoma and poorly differentiated adenocarcinoma, and this abnormality is associated with Smo protein overexpression, which upregulates the expression of the downstream transcription factor Gli1 protein.
Adenocarcinoma ; metabolism ; pathology ; physiopathology ; Adult ; Aged ; Female ; Hedgehog Proteins ; physiology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Receptors, G-Protein-Coupled ; biosynthesis ; Signal Transduction ; Smoothened Receptor ; Stomach Neoplasms ; metabolism ; pathology ; physiopathology ; Transcription Factors ; biosynthesis ; Zinc Finger Protein GLI1
9.Significance of the secreted frizzled-related protein 2 expression in earlobe keloid.
Zhi-Cheng SUN ; Chuan CAO ; Zhen-Xiang WANG ; Ju-Long WU ; Bing MA ; Shi-Rong LI
Chinese Journal of Plastic Surgery 2010;26(5):369-372
OBJECTIVETo investigate the expression and the role of secreted frizzled-related protein 2 (SFRP2) in the earlobe keloid and find a valid way to treat the keloid with gene therapy.
METHODSThe expression of SFRP2 mRNA and protein was tested with in situ hybridization and Western Blot Analysis method in the different period of earlobe keloid.
RESULTSThe SFRP2 mRNA and protein expression at the keloid edge was significantly high in 12 month group than in 3 or 6 month groups (P < 0.01), but not than in 24 month group. The SFRP2 expression started to decrease in the keloid center 12 month later (P < 0.01). The SFRP2 expression was always higher in edge than in center during all the period (P < 0.05, P < 0.01).
CONCLUSIONSThe results suggest that SFRP2 may play an important role in the development of keloid, especially at the keloid edge. The high SFRP2 expression in endothelial cells and surrounding tissue is also important. It may be a new way for gene therapy of keloid by decreasing the SFRP2 expression.
Adolescent ; Adult ; Ear ; Female ; Humans ; Keloid ; metabolism ; Male ; Membrane Proteins ; metabolism ; Middle Aged ; Young Adult
10.Hyponatremia caused by alcohol withdrawal: a case report.
Cui-xiang LIU ; Ran AO ; Bing-yuan WANG ; Da-wei XIE ; Zhen-wei WANG ; Fu-rong SUN
Chinese Journal of Hepatology 2010;18(12):948-949